The Hepatitis C Virus Non-structural NS5A Protein Impairs Both the Innate and Adaptive Hepatic Immune Response in Vivo
The Hepatitis C Virus Non-structural NS5A Protein Impairs Both the Innate and Adaptive Hepatic Immune Response in Vivo
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DOI：
10.1074/jbc.m109.038877
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发表时间：
2009-10-09
影响因子：
4.8
通讯作者：
Hildt, Eberhard
中科院分区：
文献类型：
--
作者：
Kriegs, Malte;Buerckstuemmer, Tilmann;Hildt, Eberhard
丙型肝炎病毒 (HCV) 蛋白非结构 (NS) 5A 在 HCV 相关发病机制中的作用仍然是个谜。为了研究 NS5A 对病毒持久性和病毒相关发病机制的体内作用，建立了转基因 (Tg) 小鼠模型。使用白蛋白启动子产生具有肝脏靶向 NS5A 转基因表达的小鼠。通过蛋白质印迹、淋巴细胞脉络膜脑膜炎病毒（LCMV）感染以及使用通过流体动力注射产生的瞬时NS3/4A Tg小鼠来测定肝脏免疫反应的变化。通过 Cr 释放测定法研究细胞毒性 T 淋巴细胞 (CTL) 活性。稳定的 NS5A Tg 小鼠没有表现出自发性肝病的迹象。根据体内 LCMV 感染或短暂 NS3/4A Tg 肝细胞的清除确定，NS5A Tg 小鼠的肝内免疫被破坏。这种免疫受损的原因是 LCMV 感染后干扰素 β、2',5'-OAS 和 PKR 的诱导减少以及 CTL 介导的 NS3 表达肝细胞消除受损。总之，这些数据表明在目前的转基因小鼠模型中，NS5A不会引起自发性肝病。然而，我们发现 NS5A 可能会损害先天性和适应性免疫反应，从而促进慢性 HCV 感染。
The role of hepatitis C virus (HCV) protein non-structural (NS) 5A in HCV-associated pathogenesis is still enigmatic. To investigate the in vivo role of NS5A for viral persistence and virus-associated pathogenesis a transgenic (Tg) mouse model was established. Mice with liver-targeted NS5A transgene expression were generated using the albumin promoter. Alterations in the hepatic immune response were determined by Western blot, infection by lymphocytic choriomeningitis virus (LCMV), and using transient NS3/4A Tg mice generated by hydrodynamic injection. Cytotoxic T lymphocyte (CTL) activity was investigated by the Cr-release assay. The stable NS5A Tg mice did not reveal signs of spontaneous liver disease. The intrahepatic immunity was disrupted in the NS5A Tg mice as determined by clearance of LCMV infection or transiently NS3/4A Tg hepatocytes in vivo. This impaired immunity was explained by a reduced induction of interferon beta, 2',5'-OAS, and PKR after LCMV infection and an impairment of the CTL-mediated elimination of NS3-expressing hepatocytes. In conclusion, these data indicate that in the present transgenic mouse model, NS5A does not cause spontaneous liver disease. However, we discovered that NS5A could impair both the innate and the adaptive immune response to promote chronic HCV infection.